Neuroglycan C (Chondroitin Sulfate Proteoglycan 5, Acidic Leucine-rich EGF-like Domain-containing Brain Protein, CSPG5)

Neuroglycan C (NGC; also CSPG5 and CALEB) is a 120-160kD member of the neuregulin family of proteins. Depending upon its expression profile, it may be a glycoprotein of 120kD, or a chondroitin sulfate (CS) proteoglycan of 140-160kD. NGC is expressed in nervous tissue and is found on retinal ganglion cells, cerebellar Purkinje cells and hippocampal neurons. Mature human NGC is a 536 amino acid (aa) type I transmembrane glycoprotein. It possesses a 393 aa extracellular region (aa31-423) that contains a Ser117 CS attachment site and an EGF-like domain (aa 371-413). There are at least two isoform variants. One shows a deletion of aa 487-513, while a second shows an alternaive start site at Met139 with the same deletion. Phosphorylation likely occurs on Ser249, and proteolysis generates a >75kD soluble fragment. Over aa 31-420, human NGC shares 84% aa identity with mouse NGC. Applications: Suitable for use in ELISA, Immunocytochemistry and Western Blot. Other Applications not tested. Recommended Dilution: Direct ELISA: 0.5-1ug/ml. The detection limit for recombinant human Neuroglycan C is approximately 0.3ng/well. Western Blot: 1ug/ml Immunocytochemistry: 10ug/ml was used to detect Neuroglycan C in sections of adult mouse cerebellum. Cells were fixed with PBS containing 4% paraformaldehyde for 20 minutes at room temperature and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at RT, After blocking, cells were incubated with diluted C5067-91B for 3 hours at RT followed by NorthernLights 557 donkey anti-sheep lgG at RT for 1 hour. Between each step, cells were washed with PBS containing 0.1% BSA. Optimal dilutions to be determined by the researcher. Storage and Stability: Lyophilized powder may be stored at -20 degrees C. Stable for 12 months at -20 degrees C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.